中国水稻科学2018，Vol.32Issue(2)：111-118,8.DOI:10.16819/j.1001-7216.2018.7094
水稻纹枯病菌RsPhm基因的克隆及其表达分析
Cloning and Expression Analysis of RsPhm Gene in Rhizoctonia solani AG-1ⅠA of Rice Sheath Blight Pathogen
摘要
Abstract
[Objective]In order to elucidate the functions of phenol 2-monooxygenase(RsPhm)gene in melanization of Rhizoctonia solani Kühn AG-1ⅠA,the causal agent of rice sheath blight,[Method]the gene was cloned by routine PCR and RT-PCR techniques, and the bioinformatics analysis of this gene was conducted; furthermore, the relative expression level under catechol stress was determined by using fluorescence quantitative real-time PCR (qRT-PCR) technique.[Result]Bioinformatics analysis showed that the full-length DNA and cDNA sequences of RsPhm gene were 2 628 bp and 1 983 bp,respectively,which encode 660 amino acids.The phylogenetic tree analysis showed that RsPhm gene had a close relationship in different anastomosis groups (AGs) of R.solani, and a certain evolutionary conservation among different fungal species. Results of qRT-PCR indicated that the exposure to exogenous catechol could improve the expression level of RsPhm gene,and which peaked at 12.5μg/mL of catechol,with a significant increase of 35.7 times, 19.1 times and 28.4 times up-regulated at 25 μg/mL and 50 g/mL, respectively, but only 2.1 times up-regulated at 100 g/mL.[Conclusion]The full-length sequence of RsPhm gene was obtained, its basic biological information was understood, and its expression pattern under catechol stress was clarified. These findings will lay a basis for the scientific and systematic elucidation of regulatory mechanism of melanin formation by RsPhm gene of R.solani AG-1ⅠA.关键词
水稻纹枯病菌/苯酚2-单加氧酶基因/基因克隆/表达分析Key words
Rhizoctonia solani Kühn AG-1ⅠA/RsPhm gene/gene clone/expression analysis分类
生物科学引用本文复制引用
江绍锋,王陈骄子,舒灿伟,周而勋..水稻纹枯病菌RsPhm基因的克隆及其表达分析[J].中国水稻科学,2018,32(2):111-118,8.基金项目
国家自然科学基金资助项目(31271994,31470247). （31271994,31470247）
